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首页> 外文期刊>Molecular cell >DNA Binding and ToxR Responsiveness by the Wing Domain of TcpP, an Activator of Virulence Gene Expression in Vibrio cholerae
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DNA Binding and ToxR Responsiveness by the Wing Domain of TcpP, an Activator of Virulence Gene Expression in Vibrio cholerae

机译：TcpP的翼域的DNA结合和ToxR响应性，霍乱弧菌中毒力基因表达的激活剂。

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摘要

Virulence in Vibrio cholerae requires activation of toxT by two membrane-localized activators, TcpP and ToxR. We isolated 12 tcpP activation mutants that fell into two classes: class I mutants were inactive irrespective of the presence of ToxR, and class II mutants exhibited near wild-type activity when coexpressed with ToxR. Most class I mutants had lesions in the wing domain predicted by homology with the winged helix-turn-helix family of activators. Class I mutants bound promoter DNA poorly and were largely unable to interact with ToxR in a crosslinking assay, whereas class II mutants retained physical interaction with ToxR. One mutant constructed in vitro bound DNA poorly but nevertheless responded to ToxR by activating toxT and also maintained ToxR interaction. We propose that ToxR interaction, but not DNA binding, is essential for TcP function and that the wing domain of TcpP enables contact with ToxR required for productive TcpP-RNA polymerase association.
机译：霍乱弧菌的毒力需要通过两个膜定位激活剂TcpP和ToxR激活toxT。我们分离了12个tcpP激活突变体，分为两个类别：I类突变体是无活性的，与ToxR的存在无关; II类突变体与ToxR共表达时表现出接近野生型的活性。大多数I类突变体在翼域中具有通过与翼状螺旋-转角-螺旋激活子家族同源性预测的损伤。 I类突变体与启动子DNA的结合较弱，并且在交联测定中基本无法与ToxR相互作用，而II类突变体保留了与ToxR的物理相互作用。一个突变体构建的体外结合DNA较差，但通过激活toxT仍能响应ToxR，并且还维持ToxR相互作用。我们建议ToxR相互作用，但不是DNA结合，对于TcP功能是必不可少的，并且TcpP的翼域可以与生产性TcpP-RNA聚合酶缔合所需的ToxR接触。

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